Analytical comparisons of SARS-COV-2 detection by qRT-PCR and ddPCR with multiple primer/probe sets

被引:104
作者
Liu, Xinjin [1 ]
Feng, Jiangpeng [1 ]
Zhang, Qiuhan [1 ]
Guo, Dong [1 ]
Zhang, Lu [1 ]
Suo, Tao [2 ]
Hu, Wenjia [3 ]
Guo, Ming [1 ]
Wang, Xin [1 ]
Huang, Zhixiang [1 ]
Xiong, Yong [3 ]
Chen, Guozhong [2 ]
Chen, Yu [1 ]
Lan, Ke [1 ,4 ]
机构
[1] Wuhan Univ, Coll Life Sci, Modern Virol Res Ctr, State Key Lab Virol, Wuhan, Peoples R China
[2] Wuhan Univ, Renmin Hosp, State Key Lab Virol, Wuhan, Peoples R China
[3] Wuhan Univ, Zhongnan Hosp, Dept Infect Dis, Wuhan, Peoples R China
[4] Wuhan Univ, Frontier Sci Ctr Immunol & Metab, Wuhan, Peoples R China
关键词
SARS-CoV-2; diagnosis; digital PCR; real time PCR; false positive; false negative;
D O I
10.1080/22221751.2020.1772679
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Different primers/probes sets have been developed all over the world for the nucleic acid detection of SARS-CoV-2 by quantitative real time polymerase chain reaction (qRT-PCR) as a standard method. In our recent study, we explored the feasibility of droplet digital PCR (ddPCR) for clinical SARS-CoV-2 nucleic acid detection compared with qRT-PCR using the same primer/probe sets issued by Chinese Center for Disease Control and Prevention (CDC) targeting viral ORF1ab or N gene, which showed that ddPCR could largely minimize the false negatives reports resulted by qRT-PCR [Suo T, Liu X, Feng J, et al. ddPCR: a more sensitive and accurate tool for SARS-CoV-2 detection in low viral load specimens. medRxiv [Internet]. 2020;2020.02.29.20029439. Available from: https://medrxiv.org/content/early/2020/03/06/2020.02.29.20029439.abstract]. Here, we further stringently compared the performance of qRT-PCR and ddPCR for 8 primer/probe sets with the same clinical samples and conditions. Results showed that none of 8 primer/probe sets used in qRT-PCR could significantly distinguish true negatives and positives with low viral load (10(-4) dilution). Moreover, false positive reports of qRT-PCR with UCDC-N1, N2 and CCDC-N primers/probes sets were observed. In contrast, ddPCR showed significantly better performance in general for low viral load samples compared to qRT-PCR. Remarkably, the background readouts of ddPCR are relatively lower, which could efficiently reduce the production of false positive reports.
引用
收藏
页码:1175 / 1179
页数:5
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